重组人乳头瘤病毒双价(16/18型)疫苗免疫原性评价方法的研究

Study on recombinant human papillomavirus(16/18) vaccine immunogenicity evaluation assay

目的:探讨重组人乳头瘤病毒双价(16/18型)候选疫苗免疫原性评价方法。方法:对以假病毒为基础的中和实验(Zs-Green法)测定血清中和滴度,以微球为基础建立的Luminex法测定中和滴度和ELISA法测定抗体滴度的3种方法进行相关性研究以及体内效价测定(ED50法)和体外相对效力测定(IVRP法)的相关性分析,同时对5种免疫原性的评价方法进行重复性验证。结果:ZsGreen法、Luminex法测定的中和滴度和ELISA法测定的抗体滴度检测结果表明:滴度值呈明显的剂量效应以及侯选疫苗具有良好的免疫原性。采用SPSS统计软件分析,对于HPV 16L1 ZsGreen法、Luminex法和ELISA法的相关系数分别为0.791,0.800,0.747(Spearman相关系数)和HPV 18L1的相关系数分别为0.734,0.625,0.634(Spearman相关系数)。并且ED50法和IVRP法具有一定的相关性。5种方法重复性验证的RSD分别为:5.3%(ZsGreen)、5.2%(Luminex)、8.0%(ELISA)、32.3%(ED50)、6.3%(IVRP)。结论:建立的HPV 16和HPV 18型疫苗评价方法为快速准确地评价免疫原性提供了多种解决方法。

Objective：To explore on recombinant human papillomavirus（16/18） candidate vaccine immunogenicity evaluation.Methods：To study the correlation of the ZsGreen neutralizing antibody titer（based on pseudovirus）,the average of the HPV-Luminex neutralizing antibody titer（based on beads） and the average of the ELISA antibody titer.And to explore the correlation between potency（ED50） and in vitro relative potency（IVRP）.Then we compare the repeatability validation of all five methods.Results：The results of antibody titer by ZsGreen,Luminex and ELISA show the antibody titers have obvious dose-effect relationship and candidate vaccine has good immunogenicity.By SPSS statistical analysis,ZsGreen,Luminex and ELISA correlation coefficient are 0.791,0.800,0.747（Spearman coefficient） for HPV 16L1,correlation coefficient are 0.734,0.625,0.634（Spearman coefficient） for HPV 18L1.ED50 and IVRP have certain relationship.The RSD results of five methods are 5.3%（ZsGreen）,5.2%（Luminex）,8.0%（ELISA）,32.3%（ED50）,6.3%（IVRP）.Conclusion：Established HPV（16/18） vaccine evaluation assay provides multiple solutions to evaluate vaccine immunogenicity rapidly.